肝胃同治中药复方对胆汁反流大鼠胆汁酸代谢的影响

Effect of Ganwei Tongzhi TCM Compound on bile acid metabolism of rats with bile reflux

目的观察并探讨柴胡疏肝散与旋覆代赭汤的组合中药复方对胆汁反流大鼠外周血胆汁酸代谢的影响。方法选取74只SD雄性大鼠,24只假手术组行腹腔切开和缝合术、50只进行全胃切除+食管十二指肠吻合术造模,造模成功后分为模型组和中药组。术后2周、4周、8周的模型组与假手术组使用生理盐水分别灌胃0周、2周、6周;术后8周中药组以生药量9.3 g/(kg·d)的肝胃同治复方予以灌胃,连续给药6周。各组大鼠灌胃量均为10 mL/(kg·d),经腹腔注射2%戊巴比妥钠麻醉后,开腹取食管和腹主动脉血。对食管组织进行脱水、包埋、切片和苏木素—伊红染色,观察大鼠食管组织病理学改变及癌前病变和癌变的出现率,同时用液相色谱—串联质谱法测定大鼠血清中11种胆汁酸的水平。结果术后2周到术后8周,模型组大鼠食管黏膜损伤严重程度递增,食管癌前病变和癌变的出现率逐渐上升,模型组术后8周食管癌变出现率显著高于术后2周的模型组(P<0.05);中药组大鼠食管黏膜损伤改善,食管癌变出现率与模型组相比较显著降低(P<0.05)。血清胆汁酸结果显示,与同期假手术组相比,模型组术后2周牛磺胆酸水平升高(P<0.05),牛磺鹅脱氧胆酸、脱氧胆酸水平下降(P<0.05);术后4周牛磺熊脱氧胆酸、牛磺脱氧胆酸水平升高(P<0.05),术后8周牛磺胆酸水平升高(P<0.05);与术后8周模型组比较,中药组甘氨胆酸、牛磺胆酸、牛磺鹅脱氧胆酸、牛磺脱氧胆酸水平都有所下降,但无统计学意义(P>0.05);胆酸、脱氧胆酸、熊脱氧胆酸、鹅脱氧胆酸、石胆酸、牛磺石胆酸水平有所升高,但无统计学意义(P>0.05)。结论肝胃同治中药复方具有防治胆汁反流引起食管癌变的效应,并可影响血清胆汁酸的代谢。

Objective To observe and investigate the effect of the TCM compound composed of Chaihu Shugan Powder and Xuanfu Daizhe Decoction on bile acid metabolism in peripheral blood of rats with bile reflux.Methods Among 74 SD male rats,24 in the sham group were performed laparotomy and abdominal suturing,and 50 rats were treated with complete gastrectomy and esophago-duodenostomy,which were divided into model group and TCM group after successful modeling.The model group and the sham operation group at 2,4,or 8 weeks after surgery were given intragastric administration with normal saline for 0,2,and 6 weeks,respectively.The TCM group at 8 weeks after surgery was intragastrically given with the Ganwei Tongzhi TCM Compound at a raw drug dose of 9.3 g/(kg•d)continuously for 6 weeks.The gavage volume of rats in each group was 10 mL/(kg•d).After rats anesthetized by an intraperitoneal injection of 2%pentobarbital,esophagus and the blood from abdominal aorta were collected.The esophageal tissues were dehydrated,embedded,sectioned and stained with hematoxylin-eosin.We then observed histopathological changes and the incidence of precancerous lesions and carcinogenesis of the rat esophagus,and measured the levels of 11 bile acids in rat serum by liquid chromatography-tandem mass spectrometry.Results From 2 weeks to 8 weeks after surgery,the severity of esophageal mucosa damage and the incidence of esophageal precancerous lesions and carcinogenesis of the model group were gradually increased.The incidences of esophageal carcinogenesis of the model group at 8 weeks after surgery was significantly higher than those of the model group at 2 weeks after surgery(P<0.05).The damages of esophageal mucosa of the rats in the TCM group were improved.The incidence of esophageal carcinogenesis of the TCM group was significantly lower compared with the model group(P<0.05).The results of serum bile acids showed that compared with the sham group in the same period,the taurocholic acid level was increased and the taurochenodeoxycholic acid and deoxycholic acid levels were decreased of the model group at 2 weeks after surgery(P<0.05);the tauroursodeoxycholic acid and taurodeoxycholic acid levels were increased at 4 weeks after surgery(P<0.05),and the taurocholic acid level increased at 8 weeks after surgery(P<0.05);compared with the model group at 8 weeks after surgery,the levels of glycocholic acid,taurocholic acid,taurochenodeoxycholic acid,and taurodeoxycholic acid decreased,but the difference has no statistical significant(P>0.05).And the cholic acid,deoxycholic acid,ursodeoxycholic acid,chenodeoxycholic acid,lithocholic acid,and taurolithocholic acid levels increased of the TCM group,but the difference has no statistical significant(P>0.05).Conclusion Ganwei Tongzhi TCM Compound has the effect of preventing and treating the carcinogenesis of esophagus caused by bile reflux,and can affect the metabolism of serum bile acids.